Thermostable lipases and their dynamics of improved enzymatic properties

Abstract

Thermal stability is one of the most desirable characteristics in the search for novel lipases. The search for thermophilic microorganisms for synthesising functional enzyme biocatalysts with the ability to withstand high temperature, and capacity to maintain their native state in extreme conditions opens up new opportunities for their biotechnological applications. Thermophilic organisms are one of the most favoured organisms, whose distinctive characteristics are extremely related to their cellular constituent particularly biologically active proteins. Modifications on the enzyme structure are critical in optimizing the stability of enzyme to thermophilic conditions. Thermostable lipases are one of the most favourable enzymes used in food industries, pharmaceutical field, and actively been studied as potential biocatalyst in biodiesel production and other biotechnology application. Particularly, there is a trade-off between the use of enzymes in high concentration of organic solvents and product generation. Enhancement of the enzyme stability needs to be achieved for them to maintain their enzymatic activity regardless the environment. Various approaches on protein modification applied since decades ago conveyed a better understanding on how to improve the enzymatic properties in thermophilic bacteria. In fact, preliminary approach using advanced computational analysis is practically conducted before any modification is being performed experimentally. Apart from that, isolation of novel extremozymes from various microorganisms are offering great frontier in explaining the crucial native interaction within the molecules which could help in protein engineering. In this review, the thermostability prospect of lipases and the utility of protein engineering insights into achieving functional industrial usefulness at their high temperature habitat are highlighted. Similarly, the underlying thermodynamic and structural basis that defines the forces that stabilize these thermostable lipase is discussed.

Aggregation Interface and Rigid Spots Sustain the Stable Framework of a Thermophilic N-Demethylase

“…Industrial applications almost always require stable catalysts and to promote the stabilities of enzymes is a challenge . To build a robust biocatalyst, rational design, direct evolution, and de novo design are the universal methods, via stabilizing flexible regions, introducing features from stable homologs, enhancing electrostatic interactions, and so on. , In the recent years, more attention has been paid on thermophilic microorganisms, which might produce thermostable enzymes and should be of great potential in industrial application. , In our previous work, an N -demethylase TrSOX (GenBank: ACM06094.1, PDB ID: 7EXS) with outstanding thermostability was characterized from T. roseuman extremely thermophilic microorganism in a Yellowstone hot spring, ,, together with a lipase showing marked thermostability (PDB ID: 7WOL).…”

“…25,26 In the recent years, more attention has been paid on thermophilic microorganisms, which might produce thermostable enzymes and should be of great potential in industrial application. 45,46 In our previous work, an N-demethylase TrSOX (GenBank: ACM06094.1, PDB ID: 7EXS) with outstanding thermostability was characterized from T. roseum�an extremely thermophilic microorganism in a Yellowstone hot spring, 1,19,20 together with a lipase showing marked thermostability (PDB ID: 7WOL). 21 It suggested that thousands of years' extreme environments might enforce this microorganism to produce enzymes with thermostable frameworks during natural evolution.…”

Recent Advances in the Enzymatic Synthesis of Polyester

“…In a general polymerization process, high-temperature conditions could facilitate the solubility of the substrate and shorten the reaction time while lowering reaction media viscosity. Therefore, improving the thermal stability of lipases is favorable for industrial applications, and heat-resistant lipases are one of the desirable targets for research on extreme enzymes [ 106 , 107 , 108 ]. Kamal et al [ 109 ] screened a lipase 6B from Bacillus subtilis , and it was noticed that the melting temperature (Tm, temperature at which 50% of the protein is unfolded) is 78 °C, and the optimum reaction temperature is 65 °C, respectively.…”

Combined Computer-Aided Predictors to Improve the Thermostability of Nattokinase

“…Although there have been few reports of the increased thermostability of nattokinase, the high temperature sensitivity of nattokinase limits its industrial production and application; therefore, the thermostability of nattokinase needs to be further improved. In addition, many studies have shown that there is an trade-off between the catalytic activity and the stability of enzymes; thus, the improvement of the thermostability of enzymes will inevitably reduce their catalytic activity [ 16 , 17 ]. Therefore, the design of a new nattokinase with improved thermostability and without affecting the catalytic activity has become a key issue to be solved urgently for the industrial production and oral application of nattokinase.…”